• Journal of Food Hygiene and Safety
• /
• v.6 no.3
• /
• pp.111-117
• /
• 1991

We examined to develop the enzyme linked immunosorbent assay (ELISA) for determination of zearalenone in animal feeds. Zearalenone was first converted to 6'-(carboxymethyl) zearalenone oxime(zearalenone oxime) to get a coupling site and then conjugated to bovine serum albumin(BSA) for use as immunogen and to horseradish peroxidase(HRP) for use as enzyme marker. Antibody against zearalenone was obtained after 11 weeks of immunization of rabbit with zearalenone oxime-BSA. Cross reactivity of the antibody with ${\alpha}-zearalanol,\;{\beta}-zearalenol,\;{\alpha}-zearalanol\;and\;{\beta}-zearalanol$ were 168, 46, 26 and 20% respectiviely. A simple procedure was devised for the screening of zearalenone in feeds using ELISA. Feeds samples(5g) were extracted by blending with 25 ml of methanol-phospate butTered saline-dimethylformate(70 : 29 : 1) and the extract was filtered and aqueous filterate analyzed. It took only 1 hours to do whole procedure for the analysis of zearalenone in feeds by the direct competitive ELISA, and detectable limit was 1-100 ppb. Using this procedure, only 4 of 24 feed samples showed positive results with 3.93-7.43 ppb levels.

• Food Science and Biotechnology
• /
• v.15 no.6
• /
• pp.990-992
• /
• 2006

In a previous study, we obtained polyclonal antibodies against the organophosphorus insecticide fenitrothion and developed an enzyme-linked immunosorbent assay (ELISA) for this pesticide. Using these antibodies and an enzyme tracer, a direct competitive ELISA method specific for fenitrothion using a dipstick format was developed. Dipstick ELISA using antibodies to fenitrothion immobilized on an Immunodyne membrane allowed the quick visual detection of fenitrothion at concentrations above $10\;{\mu}g/L$. The $IC_{50}$ value of dipstick ELISA using reflectance detection was $27\;{\mu}g/L$ with a detection limit of $2\;{\mu}g/L$. The recovery of fenitrothion from spiked lettuce and rice samples using the dipstick ELISA ranged from 87-107%.

• Korean Chemical Engineering Research
• /
• v.49 no.4
• /
• pp.387-392
• /
• 2011

Enzymes are being used in various fields due to their unique property of substrate specificity. Enzyme-linked immunosorbent assay(ELISA) has enabled the detection of various antigens by reporting the binding event of antigen and antibody via enzyme-catalyzed reaction. However, the sensitivity improvement of conventional ELISA has been limited because only one enzyme molecule is conjugated to one molecule of antibody. To overcome this limitation and further improve the sensitivity of ELISA, there have been efforts to increase the number ratio of enzymes to antibody. Recently, the nanobiocatalytic approaches, with their successful enzyme stabilization, improved the performance stability as well as sensitivity in a modified protocol of ELISA. The present paper introduces the basic principle of ELISA, and the recent efforts to improve sensitivity and performance stability of ELISA by using the nanobiocatalytic approaches.

• Korean Journal of Veterinary Service
• /
• v.19 no.1
• /
• pp.30-38
• /
• 1996

Enzyme-linked immunosorbent assay (ELISA) was developed to detect rotavirus antigen in fecal samples using VP6-specific monoclonal antibody(2B12). The ELISA for rotavirus antigen detection found to have specificity to all bovine and porcine rotaviruses tested but not to bovine viral diarrhea virus and bovine coronavirus. The ELISA appeared to have similar sensitivity and specificity compared to fluorescence antibody assay(FA) and electropherotyping (PAGE).

• Bulletin of the Korean Chemical Society
• /
• v.23 no.4
• /
• pp.605-609
• /
• 2002

A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of organophosphorus insecticide cyanophos. An analogue (hapten) of cyanophos was synthesized and was coupled to BSA to produce polyclonal antibodi es from rabbits. The antisera were screened against another hapten coupled to ovalbumin (OVA). Using the sera of highest specificity, an antigen-coated ELISA was developed, which showed an I50 of 310 ng/mL with the detection limit of 20 ng/mL. The antibodies showed negligible cross-reactivities with other organophosphorus pesticides except for parathion-methyl, which makes the assay suitable for the selective detection of cyanophos.

• Journal of Microbiology and Biotechnology
• /
• v.14 no.3
• /
• pp.612-619
• /
• 2004

A competitive direct enzyme-linked immunosorbent assay (cdELISA) was developed for selective and rapid detection of a glycopeptide antibiotic, teicoplanin (TP). TP was conjugated to bovine serum albumin (BSA) for use as an immunogen. Repeated subcutaneous injections of 0.5 mg of the conjugate was effective in generating specific polyclonal antibody (PAb) toward TP in rabbits, as determined by cdELISA. TP-horseradish peroxidase conjugate (TP-HRP) was used as an enzyme marker. The cdELISA was developed based on a competition reaction between TP-BSA PAb and TP-HRP conjugate. The TP-BSA PAb was highly sensitive (detection limit, 0.3 ng/ml and specific toward teicoplanin, showing no cross-reactivity to other glycopeptide antibiotics including vancomycin. There were good correlations ($r^2$=0.84 and 0.76, respectively) between cdELISA and microbiological assay, and high-performance liquid chromatography. The cdELISA system developed in this work is expected to be useful not only for selective and rapid monitoring of TP but also for study of TP pharmacokinetics.

• Food Science of Animal Resources
• /
• v.30 no.1
• /
• pp.36-41
• /
• 2010

The Hen's egg is widely used in many processed foods as an ingredient and is one of the most prevalent food allergens in children. To detect egg proteins in processed foods, we developed a competitive indirect enzyme-linked immunosorbent assay (ciELISA) using an anti-ovomucoid (OM) antibody, which was produced by immunization of rabbits with OM, the most heat-stable component of the egg proteins. The detection limit of this quantitative assay system was 30 ng/mL. Cross-reactivity of the anti-OM antibody toward OM, ovalbumin, skim milk, casein, whey protein isolate, and isolated soy protein was 100, 0.4, 0.2, 0.04, 0, and 0%, respectively. In the spike test of egg white powder in milk replacer, commercial sausage, and in-house sausage, the assay recoveries ($mean{\pm}SD$) were $129{\pm}13.7%$, $73.9{\pm}12.5%$, and $65.5{\pm}13.6%$, respectively. When egg white in a commercial crab meat analog and sausage was determined by ciELISA, the assay recovery was found to be 108% and 127%, respectively. The combined results of this study indicate that this novel ciELISA for OM detection could be applied for the quantification of hen's egg proteins in processed foods.

• Food Science of Animal Resources
• /
• v.43 no.6
• /
• pp.989-1001
• /
• 2023

Processed foods containing pork fat tissue to improve flavor and gain economic benefit may cause severe issues for Muslims, Jews, and vegetarians. This study aimed to develop an indirect enzyme-linked immunosorbent assay (iELISA) based on a monoclonal antibody specific to thermal stable-soluble protein in pork fat tissue and apply it to detect pork fat tissue in heat-processed (autoclave, steam, roast, and fry) beef meatballs. To develop a sensitive iELISA, the optimal sample pre-cooking time, coating conditions, primary and secondary dilution time, and various buffer systems were tested. The change in the iELISA sensitivity with different 96-well microtiter microplates was confirmed. The detection limit of iELISA performed with an appropriate microplate was 0.015% (w/w) pork fat in raw and heat-treated beef. No cross-reactions to other meats or fats were shown. These results mean that the iELISA can be used as an analytical method to detect trace amounts of pork fat mixed in beef.

• Journal of Microbiology and Biotechnology
• /
• v.9 no.6
• /
• pp.764-772
• /
• 1999

To develop an enzyme-linked immunosorbent assay (ELISA) for detecting mold, we produced anti-mold polyclonal antibodies by immunizing extracellular polysaccharide (EPS) of Aspergillus flavus or Penicillium citrinum in rabbits subcutaneously. Using the purified antibody (Ab) and Ab-HRP conjugate, a sandwich ELISA for EPS was established. The standard curve of the ELISA showed the detection limit for P citrinum EPS to be $0.003{\;}\mu\textrm{g}/ml$. The cross-reactivities of the anti-P citrinum EPS Ab toward components of P citrinum such as EPS, liquid, and solid culture mycelium were 100, 10.5, and 0.58%, respectively, and those toward components of A. flavus such as EPS, liquid and solid culture mycelium, and spore were 300, 0.67, 0.29, and 0%, respectively. When the reactivities toward culture broths of 59 mold strains were tested by the sandwich ELISA, most of the Aspergillus (16 of 18) and Penicillium (14 of 16) strains along with one of the two Cladosporium strains gave positive signals in the culture broths even when diluted 1,000 fold, while the rest of species such as Fusarium, Absidia, Alternaria, and Candida gave negative signals. When the water extracts of 30 corn samples were analyzed by the sandwich ELISA, the EPS in the com could be detected in the concentration range of $0.1-1.6{\;}\mu\textrm{g}/g$.

• Journal of Conservation Science
• /
• v.35 no.4
• /
• pp.363-372
• /
• 2019

Animal glue has been used as a binder in Dancheong since the Joseon dynasty. Binders play an important role in determining the physical characteristics of a painting layer. The analysis of binders can be used to identify the materials and techniques used in traditional Dancheong. Binders can be investigated using physicochemical component analyses methods such as gas chromatography/mass spectrometry, pyrolysis-gas chromatography/mass spectrometry, and fourier transform infrared spectroscopy, but the detection characteristics vary depending on the degradation properties of the pigment and binder. Therefore, cross-validation using a combination of physicochemical analysis and enzyme immunoassay is used to increase the reliability of the results. In this study, we present an enzyme-linked immunosorbent assay (ELISA) as an example of an enzyme immunoassay as a method for analyzing animal glue, a traditional binder used in Korea. The applicability of ELISA was tested using commercial animal glue, in addition to animal glue produced using a variety of extraction conditions. The animal glue was analyzed in a Noerok-additionally coated-replica sample to evaluate the possibility of analyzing the animal glue in a paint layer mixed with pigment. Based on the results, we performed an assay on the use of animal glue in the Dancheong sample of the temples of the Joseon dynasty, that are estimated to have been built in the 17^th century.